마우스 성장판 결손부에 이식된 배아기 간(幹)세포와 연골세포의 형태학적 비교

권오숙,박중규,허대영,배기원,양영철 인제대학교 2001 仁濟醫學 Vol.22 No.1

Objectives: We aimed to investigate the morphological changes of embryonic stem cells and chondrocytes. We transplanted into experimental defect of mouse growth plate. Methods and Materials: The embryonic stem cells of blastocysts were collected from superovulated mice on day 4 after the vaginal plug checked. The chondrocytes were obtained from the growth plate of the 2 weeks old mice. Embryonic stem cells and chondrocytes were transplanted into the experimental defect of the growth plate. The morphological changes of these transplanted cells were investigated under the light microscope and electron microscope. Results: Embryonic stem cells and chondrocytes began to proliferate at the 8th day and the 6th day of the culture respectively. The cell size was not decreased embryonic stem cell, but chondrocytes. The survival of embryonic stem cells began to be at the 4th day and the chondrocytes at the 8th day after transplantation on the defect. During healing process, the PCNA reaction of embryonic stem cells appeared at the 4th day, and chondrocytes at the 8th day after transplantation. The number of PCNA positive embryonic stem cells began to be increased at the 8th day, and maintained by the 20th day. The number of PCNA positive chondrocytes began to be increased at the 10th day and decreased by the 20th day. On the electron microscopic finding, the embryonic stem cells were a large euchromatic nucleus with distinct nucleolus and contained well-developed rough endoplasmic reticulum by the 10th day. At the 14th and 20th day, transplanted embryonic stem cell showed the markedly decreased cytoplasm, and had a few rough endoplasmic reticulum and small vesicles and long slender processes. Chondrocytes had well-developed rough endoplasmic reticulum by the 14th day after the transplantation. At 14th day, the long slender process of the transplanted chondrocytes separated their cytoplasm from the extracelluar matrix. Transplanted chondrocytes began to be changed into the typical chondrocytic shape with euchromatic nucleus and rich free ribosomes at the 20th day. Conclusions: The embryonic stem cell proliferated slower than the chondrocyte, but took root faster than the chondrocyte on the epiphyseal defect. Embryonic stem cells which proliferated long-standing were gradually differentiated into the chondrocytic cell and finally maintained undifferentiated cells. Chondrocytes took root slowly and were rapidly differentiated into well differentiated typical chondrocytes.

Bone Formation of Embryonic Stem Cell-Derived Mesenchymal Stem Cells

정연태,유기연,이희수 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.4

Human embryonic stem cells are multipotent cells. In this study, We observed osteogenesis of human embryonic stem cell derived mesenchymal stem cells. mbryonic body formation method was used to derive mesenchymal stem cells from human embryonic stem cells. Embryonic stem cell derived mesenchymal stem cells were immunostained for CD 73 to make characterization of mesencymal stem cells. Osteogenesis of CD 73 positive mesencymal stem cells with media included ascorbic acid, dexamethasone and glycerophosphate was induced. After 10 days culture with osteogenesis media, the cells were immunostained with type I collagen, osteocalcin and Runx-2. After 21 days culture with osteogenesis media, the cells were stained with alizarin red to observe bone nodule formation. Mesenchymal stem cells were derived from human embryonic stem cells by embryonic body method. After these cells were cultured with osteogenesis media, the cells were differentiated into osteoblast and showed bone nodule formation.

Bone Formation of Embryonic Stem Cell-Derived Mesenchymal Stem Cells

( Yeon Tae Jung ),( Ki Yeon Yoo ),( Hee Su Lee ) 한국조직공학·재생의학회 2015 조직공학과 재생의학 Vol.12 No.2s

Human embryonic stem cells are multipotent cells. In this study, We observed osteogenesis of human embryonic stem cell derived mesenchymal stem cells. mbryonic body formation method was used to derive mesenchymal stem cells from human embryonic stem cells. Embryonic stem cell derived mesenchymal stem cells were immunostained for CD 73 to make characterization of mesencymal stem cells. Osteogenesis of CD 73 positive mesencymal stem cells with media included ascorbic acid, dexamethasone and glycerophosphate was induced. After 10 days culture with osteogenesis media, the cells were immunostained with type I collagen, osteocalcin and Runx-2. After 21 days culture with osteogenesis media, the cells were stained with alizarin red to observe bone nodule formation. Mesenchymal stem cells were derived from human embryonic stem cells by embryonic body method. After these cells were cultured with osteogenesis media, the cells were differentiated into osteoblast and showed bone nodule formation. Tissue Eng Regen Med 2015;12(Suppl 2):132-137

인간배아연구윤리강령 개발에 관한 연구

이은영(Lee Eun Young) 한국생명윤리학회 2005 생명윤리 Vol.6 No.1

The South Korean investigators and scientists created embryos that were genetic copies of women who donated their cells. After a few days, some of the embryos had grown to a stage when embryonic stem cells could be extracted, and the scientists created a culture line of such a stem cell. The stem cells have the ability to turn into any other kinds of cells in the body. At the blastocyst stage, the inner cell mass consists of cells that are still indeterminate, not yet committed to becoming any particular cell type. They are the stem cells, which can in theory develop into any of the body's tissues and organs. Stem cells from a clone would be genetically identical to the person who contributed cells to make the embryo. Some researchers believe stem cells may hold the key to curing diseases such as diabetes, Parkinson's and Alzheimer's. Doctors want to be able to repair damaged tissues with a patient's own genetic material. Some scientists say it would be more practical to use stem cells from adults. While some experts say these cells cannot be grown outside the body as easily as embryonic stem cells and may not be as versatile, they are more predictable in what kind of cells they turn into. Use of adult stem cells is not ethically objectionable. But embryonic stem cells are ethically objectionable. I will point out ethical problems of using embryonic stem cells, and make a counterproposal of theses problems. This paper undertakes to provide a philosophical grounding for ethically responsible research of embryonic stem cells. As a result, I'll maintain that researchers of embryonic stem cell must develop and implement codes of research ethics in accordance with the public opinion. This paper will go far toward solving the ethical problem of donating and obtaining the human ova.

인간배아줄기세포연구의 민사법적 의미

김민중(KIM Min-Joong) 한국법학원 2008 저스티스 Vol.- No.103

인간배아줄기세포연구에 대한 사회적 논쟁이 뜨겁다. 인간배아줄기세포연구가 척수장애, 파킨슨, 알츠하이머, 뇌졸중, 당뇨와 같은 난치병을 퇴치할 수 있다고 하는 의미에서는 이보다 인류에 더 공헌할 수 있는 일을 찾기 어렵다고 할 수 있다. 그러나 인간배아줄기세포연구가 사회적, 윤리적, 법적으로 아무런 문제점도 없다고 할 수는 없다. 인간배아줄기세포의 연구나 이용과 관련하여 선결하여야 할 과제도 적지 않고, 인간배아줄기세포연구에는 과학적인 문제뿐만 아니라 여러 가지 해결하여야 할 윤리적, 법적 문제가 내포되어 있다는 사실을 부인할 수 없다. 현재 인간배아줄기세포의 연구나 이용에 관한 생명윤리의 문제는 국내에서 뿐만 아니라, 전세계적으로도 대단한 논란이 되고 있다. 또한 인간배아줄기세포연구가 야기하는 문제는 생명윤리의 문제에 국한되지 아니한다. 법적으로도 인간배아줄기세포연구와 관련한 다양한 문제가 제기된다. 인간배아줄기세포연구에 관한 근본적인 법적 문제로서는 인간의 존엄성와 관련한 인권문제가 제기된다. 그리고 예를 들어 인간배아 줄기세포연구를 형법적으로 금지하여야 하는가 하는 형법상의 문제도 제기된다. 역시 인간배아줄기세포연구에서의 법적 문제에 관한 핵심적인 내용의 하나는 인간배아줄기세포연구를 둘러싼 민사법적 과제라고 할 수 있다. 인간배아줄기세포연구와 관련하여 제기되는 민사법적 과제는 매우 다양하리라고 예상되며, 계약법적 문제를 비롯하여 책임법적 문제, 가족법적 문제에까지 민법 전반에 걸쳐 문제가 제기될 수 있다. 우선 인간배아줄기세포연구와 관련하여 다양한 계약관계가 성립할 수 있다. 예를 들어 인간배아줄기세포연구를 위하여는 배아, 특히 난자가 필요하므로, 배아나 난자를 제공받기 위하여는 배아제공자 및 난자 제공자와의 사이에 배아제공계약 또는 난자제공계약이 성립한다. 또한 줄기세포를 난치성 질환을 치료하기 위한 세포치료의 재료로 사용하는 경우에는 환자와의 사이에 첨단의료행위로서의 임상실험이나 줄기세포치료를 실시하기 위한 임상실험계약 또는 의료계약이 성립한다. 인간배아줄기세포연구와 관련한 임상실험을 통하여 피실험자나 제3자가 손해를 입을 수 있다. 만약 줄기세포연구를 적용한 임상실험에서 피실험자나 제3자에게 손해가 발생하면 손해배상책임이 문제되고, 보통 임상실험으로 생긴 손해에 따른 피실험자에 대한 책임은 계약책임이 되고, 제3자에 대한 책임은 불법행위책임으로 된다. The debate over the treatment of human stem cell is new. Ever since human stem cells were first isolated, the possible applications of stem cell research and the moral and legal issues surrounding human stem cell research have created much controversy. Now human stem cell research is a controversial and divisive topic. Research on embryonic stem cells has generated great intrigue in the scientific community. Many medical researchers consider stem cell-based therapies to have the potential of treating a host of human illnesses and yielding a number of medical benefits. However, the embryonic stem cell research raises numerous ethical and legal concerns. The embryonic stem cell research destroys the human embryo. This has generated a storm of debate about if this research can be legally and ethically justified. Human embryonic stem cell is 'master cell', able to develop into almost any cell in the human body. The research with human embryonic stem cells raises complex and sensitive legal questions that should be considered carefully and discussed widely. The civil legal debate surrounding human embryonic stem cell research, will be the focus of this paper.

Pig Pluripotent Stem Cells as a Candidate for Biomedical Application

Kwang-Hwan Choi,Chang-Kyu Lee 한국동물생명공학회(구 한국동물번식학회) 2019 Journal of Animal Reproduction and Biotechnology Vol.34 No.3

Stem cells are progenitor cells that are capable of self-renewal and differentiation into various cells. Especially, pluripotent stem cells (PSCs) have in vivo and in vitro differentiation capacity into three germ layers and can proliferate infinitely. The differentiation ability of PSCs can be applied for regenerative medicine and tissue engineering. In domestic animals, their PSCs have a potential for preclinical therapy as well as the production of transgenic animals and agricultural usage such as cultured meat. Among several domestic animals, a pig is considered as an ideal model for biomedical and agricultural purposes mentioned above. In this reason, studies for pig PSCs including embryonic stem cells (ESCs), embryonic germ cells (EGCs) and induced pluripotent stem cells (iPSCs) have been conducted for decades. Therefore, this review will discuss the history of PSCs derived from various origins and recent progress in pig PSC research field.

Expression Profile of the Stem Cell Markers in Human Hertwig’s Epithelial Root Sheath/Epithelial Rests of Malassez Cells

남현,이진,Jaewon Kim,박재완,박주철,김정욱,서병무,Jae Cheoun Lee 한국분자세포생물학회 2011 Molecules and cells Vol.31 No.4

Hertwig’s epithelial root sheath/Epithelial rests of Malassez (HERS/ERM) cells are unique epithelial cells in the periodontal ligament. They remain in periodontal tissues throughout the adult life, and it is expected that their functional role is to maintain the homeostasis of the periodontium through reciprocal interactions with other periodontal cells. In this study, we investigated whether HERS/ERM cells have primitive stem cell characteristics: those of embryonic stem cells as well as of epithelial stem cells. Primary HERS/ERM cells had typical epithelial cell morphology and characteristics and they maintained for more than five passages. They expressed epithelial stem cell-related genes: ABCG2, ΔNp63, p75, EpCAM, and Bmi-1. Moreover,the expression of embryonic stem cell markers such as Oct-4, Nanog, and SSEA-4 were detected. Next, we investigated whether the expression of these stem cell markers was maintained during the sub-culture process. HERS/ERM cells showed different expression levels of these stemness genes at each passage, but their expression was maintained throughout the passages. Taken together,our data suggest that a primary culture of HERS/ERM cells contains a population of primitive stem cells that express epithelial stem cell markers and embryonic stem cell markers. Furthermore, these cell populations were maintained during the sub-culturing process in our culture conditions. Therefore, our findings suggest that there is a strong possibility of accomplishing cementum tissue engineering with HERS/ERM cells.

Profiling of Differentially Expressed Genes in Human Stem Cells by cDNA Microarray

김철근,이종주,정대영,전진선,허현석,강호철,신준호,조윤신,차경준,김찬길,도병록,김경숙,김현수 한국분자세포생물학회 2006 Molecules and cells Vol.21 No.3

Stem cells are unique cell populations with the ability to undergo both self-renewal and differentiation, although a wide variety of adult stem cells as well as embryonic stem cells have been identified and stem cell plasticity has recently been reported. To identify genes implicated in the control of the stem cell state as well as the characteristics of each stem cell line, we analyzed the expression profiles of genes in human embryonic, hematopoietic (CD34+ and CD133+), and mesenchymal stem cells using cDNA microarrays, and identified genes that were differentially expressed in specific stem cell populations. In particular we were able to identify potential hESC signature-like genes that encode transcription factors (TFAP2C and MYCN), an RNA binding protein (IMP-3), and a functionally uncharacterized protein (MAGEA4). The overlapping sets of 22 up-regulated and 141 downregulated genes identified in this study of three human stem cell types may also provide insight into the developmental mechanisms common to all human stem cells. Furthermore, our comprehensive analyses of gene expression profiles in various adult stem cells may help to identify the genetic pathways involved in self-renewal as well as in multi-lineage specific differentiation.

배아줄기세포

황정혜 한양대학교 의과대학 2002 한양의대 학술지 Vol.22 No.2

Stem cells are unique cell populations with the ability to undergo both self-renewal and differentiation. A wide variety of adult mammalian tissues harbors stem cells, yet "adult" stem cells may be capable of developing into only a limited number of cell types. In contrast, embryonic stem (ES) cells, derived from blastocyst-stage early mammalian embryos, have the ability to form any fully differentiated cell of the body. Human ES cells have a normal karyotype, maintain high telomerase activity, and exhibit remarkable long-term proliferative potential, providing the possibility for unlimited expansion in culture. Furthermore, they can differentiate into derivatives of all three embryonic germ layers when transferred to an in vivo environment. Because they have the dual ability to proliferate indefinitely and differentiate into multiple tissue types, human ES cells could potentially provide an unlimited supply of tissue for human transplantation. Though human ES cell-based transplantation therapy holds great promise to successfully treat a variety of diseases(e.g., Parkinson's disease, diabetes, and heart failure) many barriers remain in the way of successful clinical trials. This review summarize the current status of stem cell research using pluripotent stem cells in the areas such as derivation, establishment and characterization of the stem cell lines, regulatory factors during differentiation, and ethical problems in stem cell research.

Stem cell therapy in pain medicine

Yong Hee Han,Kyung Hoon Kim,Salahadin Abdi,Tae Kyun Kim 대한통증학회 2019 The Korean Journal of Pain Vol.32 No.4

Stem cells are attracting attention as a key element in future medicine, satisfying the desire to live a healthier life with the possibility that they can regenerate tissue damaged or degenerated by disease or aging. Stem cells are defined as undifferentiated cells that have the ability to replicate and differentiate themselves into various tissues cells. Stem cells, commonly encountered in clinical or preclinical stages, are largely classified into embryonic, adult, and induced pluripotent stem cells. Recently, stem cell transplantation has been frequently applied to the treatment of pain as an alternative or promising approach for the treatment of severe osteoarthritis, neuropathic pain, and intractable musculoskeletal pain which do not respond to conventional medicine. The main idea of applying stem cells to neuropathic pain is based on the ability of stem cells to release neurotrophic factors, along with providing a cellular source for replacing the injured neural cells, making them ideal candidates for modulating and possibly reversing intractable neuropathic pain. Even though various differentiation capacities of stem cells are reported, there is not enough knowledge and technique to control the differentiation into desired tissues in vivo. Even though the use of stem cells is still in the very early stages of clinical use and raises complicated ethical problems, the future of stem cells therapies is very bright with the help of accumulating evidence and technology.